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      Alexa Fluor™ 647 Mouse Anti-Human IL-12/IL-23 p40
      Alexa Fluor™ 647 Mouse Anti-Human IL-12/IL-23 p40
      Two-color flow cytometric analysis of IL-12/IL-23 p40 expression in stimulated Human monocytes.  Human peripheral blood mononuclear cells (PBMC) were preincubated (2 hr) with Recombinant Human IFN-γ (Cat. No. 554617/554616; 10 ng/ml). Lipopolysaccharide (LPS; Sigma, Cat. No. L-2654; 1.0 μg/ml) and BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724] were then added to the IFN-γ-primed cells which were further cultured overnight. The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 555397) and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 568947; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human IL-12/IL-23 p40 antibody (Cat. No. 570331/570407; Right Plot) using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-12/IL-23 p40 (or Ig Isotype control staining) versus CD14 was derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      Alexa Fluor™ 647 Mouse Anti-Human IL-12/IL-23 p40
      Two-color flow cytometric analysis of IL-12/IL-23 p40 expression in stimulated Human monocytes.  Human peripheral blood mononuclear cells (PBMC) were preincubated (2 hr) with Recombinant Human IFN-γ (Cat. No. 554617/554616; 10 ng/ml). Lipopolysaccharide (LPS; Sigma, Cat. No. L-2654; 1.0 μg/ml) and BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724] were then added to the IFN-γ-primed cells which were further cultured overnight. The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 555397) and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 568947; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human IL-12/IL-23 p40 antibody (Cat. No. 570331/570407; Right Plot) using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-12/IL-23 p40 (or Ig Isotype control staining) versus CD14 was derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      Product Details
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      BD Pharmingen™
      IL12B; IL-12B; IL12p40; IL-12-40; CLMF; CLMF2; NKSF; NKSF2
      Human (QC Testing)
      Mouse IgG1, κ
      Purified p40 Subunit of Human IL-12
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl/test
      3593
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.  However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      11. For U.S. patents that may apply, see bd.com/patents.
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      570407 Rev. 1
      Antibody Details
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      C8.6

      The C8.6 monoclonal antibody specifically recognizes human IL-12 p40 that is encoded by IL12B (interleukin 12B). The antibody binds to IL-12 p40 monomers and homodimers as well as to the p40 subunit of the heterodimeric cytokines, IL-12 p70 (p35/p40) and IL-23 (p19/p40). These cytokines are produced by a variety of cells, including activated monocytes, macrophages, neutrophils, and dendritic cells. IL-12 p40 monomers and homodimers can block the binding of heterodimeric IL-12 to its receptor.  IL-12 promotes the generation of Th1 cells and activates NK cells, Group 1 innate lymphoid cells (ILC1), and cytotoxic T cells leading to production of IFN-γ, TNF, granzymes, and perforin. IL-23 promotes the generation and activation of Th17 cells and other effector cells, including γδ T cells, NKT cells, MAIT cells, and ILC3 that can produce cytokines including IL-17A, IL-17F, IL-22, GM-CSF, TNF, and IL-6. The immunogen used to generate the C8.6 hybridoma was the purified p40 subunit of human IL-12. The C8.6 antibody is a neutralizing antibody and can reportedly inhibit the biological activities of IL-12 and IL-23.

      570407 Rev. 1
      Format Details
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      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      570407 Rev.1
      Citations & References
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      View product citations for antibody "570407" on CiteAb

      Development References (4)

      1. D'Andrea A, Aste-Amezaga M, Valiante NM, Ma X, Kubin M, Trinchieri G. Interleukin 10 (IL-10) inhibits human lymphocyte interferon gamma-production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells. J Exp Med. 1993; 178(3):1041-1048. (Clone-specific: Neutralization). View Reference
      2. D'Andrea A, Rengaraju M, Valiante NM, et al. Production of natural killer cell stimulatory factor (interleukin 12) by peripheral blood mononuclear cells. J Exp Med. 1992; 176(5):1387-1398. (Clone-specific: Neutralization, Radioimmunoassay, Western blot). View Reference
      3. Gerosa F, Baldani-Guerra B, Lyakh LA, et al. Differential regulation of interleukin 12 and interleukin 23 production in human dendritic cells.. J Exp Med. 2008; 205(6):1447-61. (Clone-specific: Functional assay, Immunoprecipitation, Neutralization). View Reference
      4. Oppmann B, Lesley R, Blom B, et al. Novel p19 protein engages IL-12p40 to form a cytokine, IL-23, with biological activities similar as well as distinct from IL-12.. Immunity. 2000; 13(5):715-25. (Clone-specific: Immunoprecipitation, Neutralization). View Reference
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      570407 Rev. 1

       

      Please refer to Support Documents for Quality Certificates


      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

      For Research Use Only. Not for use in diagnostic or therapeutic procedures.